中国医药生物技术
中國醫藥生物技術
중국의약생물기술
CHINESE MEDICINAL BIOTECHNOLOGY
2014年
5期
347-352
,共6页
韩芳%陆巧妮%徐力昆%窦媛媛%李玉环
韓芳%陸巧妮%徐力昆%竇媛媛%李玉環
한방%륙교니%서력곤%두원원%리옥배
抗病毒药%人乳头瘤病毒 16%宫颈肿瘤%西多福韦
抗病毒藥%人乳頭瘤病毒 16%宮頸腫瘤%西多福韋
항병독약%인유두류병독 16%궁경종류%서다복위
Antiviral agents%Human papillomavirus 16%Uterine cervical neoplasms%Cidofovir
目的从抗病毒角度探讨西多福韦对宫颈癌细胞 CaSki内人乳头瘤病毒16（HPV16）的抑制作用及对细胞周期的影响。方法用 MTT法检测西多福韦对细胞的毒性；用实时定量 PCR法检测其对病毒 E6、E7 mRNA水平的影响；用 Western blot方法检测其对病毒蛋白 E6、E7和细胞抑癌蛋白 p53、pRb表达水平的影响；用流式细胞法检测其对宫颈癌细胞周期的影响。结果西多福韦对宫颈癌细胞毒性较正常细胞大。可使HPV16阳性宫颈癌细胞 CaSki内 E6、E7 mRNA和蛋白水平降低，最大抑制率分别为（33.38±8.00）%、（28.32±2.73）%和98.92%、97.46%；可以使 p53、pRb蛋白水平升高，最大浓度时可以上调12.06和3.53倍；对 HPV16阴性宫颈癌细胞 C-33A p53蛋白表达无影响，但可提高 pRb蛋白水平；可导致 CaSki和 C-33A细胞发生 S期阻滞，最高浓度组细胞相对对照组 S期分别增加22.83%和67.64%。结论西多福韦可以在对细胞无毒的浓度下，抑制宫颈癌细胞内的 HPV16，诱导宫颈癌细胞发生 S期阻滞。
目的從抗病毒角度探討西多福韋對宮頸癌細胞 CaSki內人乳頭瘤病毒16（HPV16）的抑製作用及對細胞週期的影響。方法用 MTT法檢測西多福韋對細胞的毒性；用實時定量 PCR法檢測其對病毒 E6、E7 mRNA水平的影響；用 Western blot方法檢測其對病毒蛋白 E6、E7和細胞抑癌蛋白 p53、pRb錶達水平的影響；用流式細胞法檢測其對宮頸癌細胞週期的影響。結果西多福韋對宮頸癌細胞毒性較正常細胞大。可使HPV16暘性宮頸癌細胞 CaSki內 E6、E7 mRNA和蛋白水平降低，最大抑製率分彆為（33.38±8.00）%、（28.32±2.73）%和98.92%、97.46%；可以使 p53、pRb蛋白水平升高，最大濃度時可以上調12.06和3.53倍；對 HPV16陰性宮頸癌細胞 C-33A p53蛋白錶達無影響，但可提高 pRb蛋白水平；可導緻 CaSki和 C-33A細胞髮生 S期阻滯，最高濃度組細胞相對對照組 S期分彆增加22.83%和67.64%。結論西多福韋可以在對細胞無毒的濃度下，抑製宮頸癌細胞內的 HPV16，誘導宮頸癌細胞髮生 S期阻滯。
목적종항병독각도탐토서다복위대궁경암세포 CaSki내인유두류병독16（HPV16）적억제작용급대세포주기적영향。방법용 MTT법검측서다복위대세포적독성；용실시정량 PCR법검측기대병독 E6、E7 mRNA수평적영향；용 Western blot방법검측기대병독단백 E6、E7화세포억암단백 p53、pRb표체수평적영향；용류식세포법검측기대궁경암세포주기적영향。결과서다복위대궁경암세포독성교정상세포대。가사HPV16양성궁경암세포 CaSki내 E6、E7 mRNA화단백수평강저，최대억제솔분별위（33.38±8.00）%、（28.32±2.73）%화98.92%、97.46%；가이사 p53、pRb단백수평승고，최대농도시가이상조12.06화3.53배；대 HPV16음성궁경암세포 C-33A p53단백표체무영향，단가제고 pRb단백수평；가도치 CaSki화 C-33A세포발생 S기조체，최고농도조세포상대대조조 S기분별증가22.83%화67.64%。결론서다복위가이재대세포무독적농도하，억제궁경암세포내적 HPV16，유도궁경암세포발생 S기조체。
Objective To investigate the antiviral activity of cidofovir (CDV) against HPV type 16 in the CaSki cervical cancer cell line and its effects on the cell cycle. Methods Cytotoxicities of CDV in CaSki, C-33A and HEL were assessed by MTT assay. The mRNA and protein levels of E6 and E7 oncogene were analyzed by quantitative real-time PCR (qRT-PCR) and Western blot. p53 and pRb protein levels were also detected by Western blot. The effect of CDV on cell cycle was analyzed by flow cytometry. Results MTT assay showed that cytotoxicity of CDV was much greater in cervical carcinoma cells than in normal cells. In HPV16-positive CaSki cervical carcinoma cell line, qRT-PCR results showed that E6 and E7 mRNA levels were decreased. Meanwhile, Western blot analysis revealed that E6 and E7 protein levels had the same trend with mRNA. However, p53 and pRb protein expressions were found to be increased simultaneously. Moreover, pRb protein expression was found to be increased in the HPV16-negative C-33A cervical carcinoma cell line, while the level of p53 protein didn’t change. In addition, flow cytometry assay showed CDV could induce S phase arrest in both cervical carcinoma cell lines. Conclusion This study showed that cidofovir has antiviral activity through suppressing E6 and E7 oncogene expressions and could induce S phase arrest at nontoxic concentration.
